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In this article, we demonstrate a plasmo-thermal bacterial accumulation effect using a miniature plasmonic optical fiber. The combined action of far-field convection and a near-field trapping force (referred to as thermophoresis)-induced by highly localized plasmonic heating-enabled the large-area accumulation of Escherichia coli. The estimated thermophoretic trapping force agreed with previous reports, and we applied speckle imaging analysis to map the in-plane bacterial velocities over large areas. This is the first time that spatial mapping of bacterial velocities has been achieved in this setting. Thus, this analysis technique provides opportunities to better understand this phenomenon and to drive it towards in vivo applications.Photobiomodulation therapy (PBMT) uses light to stimulate cells. The molecular basis of the effects of PBMT is being unveiled, but it is stated that the cytochrome-c oxidase enzyme in mitochondria, a photon acceptor of PBMT, contributes to an increase in ATP production and modulates the reduction and oxidation of electron carriers NADH and FAD. Since its effects are not fully understood, PBMT is not used on tumors. Thus, it is interesting to investigate if its effects correlate to mitochondrial metabolism and if so, how it could be linked to the optical redox ratio (ORR), defined as the ratio of FAD/(NADH + FAD) fluorescences. To that end, fibroblasts (HDFn cell line) and oral squamous cell carcinoma (SCC-25 cell line) were irradiated with a light source of 780 nm and a total dose of 5 J/cm2, and imaged by optical microscopy. PBMT down-regulated the SCC-25 ORR by 10%. Furthermore, PBMT led to an increase in ROS and ATP production in carcinoma cells after 4 h, while fibroblasts only had a modest ATP increase 6 h after irradiation. Cell lines did not show distinct cell cycle profiles, as both had an increase in G2/M cells. This study indicates that PBMT decreases the redox state of oral cancer by possibly increasing glycolysis and affects normal and tumor cells through distinct pathways. To our knowledge, this is the first study that investigated the effects of PBMT on mitochondrial metabolism from the initiation of the cascade to DNA replication. This is an essential step in the investigation of the mechanism of action of PBMT in an effort to avoid misinterpretations of a variety of combined protocols.Light-field fluorescence microscopy can record large-scale population activity of neurons expressing genetically-encoded fluorescent indicators within volumes of tissue. Conventional light-field microscopy (LFM) suffers from poor lateral resolution when using wide-field illumination. Here, we demonstrate a structured-illumination light-field microscopy (SI-LFM) modality that enhances spatial resolution over the imaging volume. This modality increases resolution by illuminating sample volume with grating patterns that are invariant over the axial direction. The size of the SI-LFM point-spread-function (PSF) was approximately half the size of the conventional LFM PSF when imaging fluorescent beads. SI-LFM also resolved fine spatial features in lens tissue samples and fixed mouse retina samples. Finally, SI-LFM reported neural activity with approximately three times the signal-to-noise ratio of conventional LFM when imaging live zebrafish expressing a genetically encoded calcium sensor.Cancer metastasis after traditional surgery introduces a high barrier to therapy efficacy. Photodynamic therapy (PDT) for cancer is based on a photochemical process of photosensitizers that concentrate in tumors and release oxidant species under light excitation to destroy cells. Compared with traditional surgery, PDT provides minimal invasion and targeted therapy. In this in vivo study, we monitor the real-time and long-term dynamics of circulating tumor cells (CTCs) after a single round of PDT and after surgical resection in a breast cancer animal model. The CTC level is low after PDT treatment, and the recurrence of the primary tumor is postponed in the PDT group compared with the resection group. We find that metastasis is correlated with the CTC level, and the PDT-treated mice show no metastasis in the lung or liver. Our results suggest PDT can effectively reduce metastasis by minimizing CTCs after treatment and is a great technology for breast cancer therapy.We performed full circumferential imaging of the Schlemm's canal (SC) of two human eyes using a Fourier domain mode-lock laser (FDML) based 1.66-MHz SS-OCT prototype at 1060 nm. Eight volumes with overlapping margins were acquired around the limbal area with customized raster scanning patterns designed to fully cover the SC while minimizing motion artifacts. The SC was segmented from the volumes using a semi-automated active contour segmentation algorithm, whose mean dice similarity coefficient was 0.76 compared to the manual segmentation results. We also reconstructed three-dimensional (3D) renderings of the 360° SC by stitching the segmented SCs from the volumetric datasets. Quantitative metrics of the full circumferential SC were provided, including the mean and standard deviation (SD) of the cross-sectional area (CSA), the maximum CSA, the minimum and maximum SC opening width, and the number of collector channels (CC) stemming from the SC.We demonstrate computational multi-directional optical coherence tomography (OCT) to assess the directional property of tissue microstructure. This method is the combination of phase-sensitive volumetric OCT imaging and post-signal processing. The latter comprises of two steps. The first step is an intensity-directional analysis, which determines the dominant en face fiber orientations. The second step is the phase-directional imaging, which reveals the sub-resolution depth-orientation of the microstructure. The feasibility of the method was tested by assessing muscle and tendon samples. Stripe patterns with several sizes were visualized in the phase-directional images. In order to interpret these images, the muscle and tendon structures were numerically modeled, and the phase-directional images were generated from the numerical model. The similarity of the experimental and numerical results suggested that the stripe patterns correspond to the muscle fiber bundle and its crimping.The adaptive matched filter (AMF) is a method widely used in spectral unmixing to classify different tissue chromophores in photoacoustic images. this website However, a threshold needs to be applied to the AMF detection image to distinguish the desired tissue chromophores from the background. link2 In this study, we propose an automatic threshold selection (ATS) algorithm capable of differentiating a target from the background, based on the features of the AMF detection image. The mean difference between the estimated thickness, using the ATS algorithm, and the known values was 0.17 SD (0.24) mm for the phantom inclusions and -0.05 SD (0.21) mm for the tissue samples of malignant melanoma. The evaluation shows that the thickness and the width of the phantom inclusions and the tumors can be estimated using AMF in an automatic way after applying the ATS algorithm.A theoretical method to determine the optimum laser parameters for maximizing the cutting efficiency for different materials (in particular human cornea) is proposed. The model is simple and reduced to laser beam characteristics and cavitation properties. The model further provides a method to convert energy fluctuations during the cutting process to equivalent deviations in the cavitation bubbles. The proposed model can be used for calibration, verification and validation purposes of laser systems used for cutting processes at relatively low cost and may improve the quality of the results.Conventional fluorescence molecular tomography (FMT) reconstruction requires photons penetrating the whole object, which limits its applications to small animals. However, by utilizing reflective photons, fluorescence distribution near the surface could be reconstructed regardless of the object size, which may extend the applications of FMT to surgical navigation and so on. Therefore, time-domain reflective fluorescence molecular tomography (TD-rFMT) is proposed in this paper. The system excites and detects the emission light from the same angle within a field of view of 5 cm. Because the detected intensities of targets depend strongly on the depth, the reconstruction of targets in deep regions would be evidently affected. Therefore, a fluorescence yield reconstruction method with depth regularization and a weighted separation reconstruction strategy for lifetime are developed to enhance the performance for deep targets. Through simulations and phantom experiments, TD-rFMT is proved capable of reconstructing fluorescence distribution within a 2.5-cm depth with accurate reconstructed yield, lifetime, and target position(s).Patient-derived cancer organoids (PCOs) are in vitro organotypic models that reflect in vivo drug response, thus PCOs are an accessible model for cancer drug screening in a clinically relevant timeframe. However, current methods to assess the response of PCOs are limited. Here, a custom swept-source optical coherence tomography (OCT) system was used to rapidly evaluate volumetric growth and drug response in PCOs. This system was optimized for an inverted imaging geometry to enable high-throughput imaging of PCOs. An automated image analysis framework was developed to perform 3D single-organoid tracking of PCOs across multiple time points over 48 hours. Metabolic inhibitors and cancer therapies decreased PCOs volumetric growth rate compared to control PCOs. Single-organoid tracking improved sensitivity to drug treatment compared to a pooled analysis of changes in organoid volume. OCT provided a more accurate assessment of organoid volume compared to a volume estimation method based on 2D projections. Single-organoid tracking with OCT also identified heterogeneity in drug response between solid and hollow PCOs. This work demonstrates that OCT and 3D single-organoid tracking are attractive tools to monitor volumetric growth and drug response in PCOs, providing rapid, non-destructive methods to quantify heterogeneity in PCOs.A deeper understanding of spatial resolution has led to innovations in microscopy and the disruption of biomedical research, as with super-resolution microscopy. To foster similar advances in time-resolved and spectral imaging, we have previously introduced the concept of 'biochemical resolving power' in fluorescence microscopy. link3 Here, we apply those concepts to investigate how the instrument response function (IRF), sampling conditions, and photon-statistics limit the biochemical resolution of fluorescence lifetime microscopy. Using Fisher information analysis and Monte Carlo simulations, we reveal the complex dependencies between photon-statistics and the IRF, permitting us to quantify resolution limits that have been poorly understood (e.g., the minimum resolvable decay time for a given width of the IRF and photon-statistics) or previously underappreciated (e.g., optimization of the IRF for biochemical detection). With this work, we unravel common misunderstandings on the role of the IRF and provide theoretical insights with significant practical implications on the design and use of time-resolved instrumentation.

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